Department of Chemical and Biomolecular Engineering, Samueli School of Engineering, University of California, Irvine.
Department of Biomedical Engineering, Samueli School of Engineering, University of California, Irvine.
J Vis Exp. 2023 Jun 16(196). doi: 10.3791/64909.
Human mesenchymal stem cells (hMSCs) offer a patient-derived cell source for conducting mechanistic studies of diseases or for several therapeutic applications. Understanding hMSC properties, such as their electrical behavior at various maturation stages, has become more important in recent years. Dielectrophoresis (DEP) is a method that can manipulate cells in a nonuniform electric field, through which information can be obtained about the electrical properties of the cells, such as the cell membrane capacitance and permittivity. Traditional modes of DEP use metal electrodes, such as three-dimensional electrodes, to characterize the response of cells to DEP. In this paper, we present a microfluidic device built with a photoconductive layer capable of manipulating cells through light projections that act as in situ virtual electrodes with readily conformable geometries. A protocol is presented here that demonstrates this phenomenon, called light-induced DEP (LiDEP), for characterizing hMSCs. We show that LiDEP-induced cell responses, measured as cell velocities, can be optimized by varying parameters such as the input voltage, the wavelength ranges of the light projections, and the intensity of the light source. In the future, we envision that this platform could pave the way for technologies that are label-free and perform real-time characterization of heterogeneous populations of hMSCs or other stem cell lines.
人骨髓间充质干细胞 (hMSCs) 为开展疾病的机制研究或多种治疗应用提供了一种源自患者的细胞来源。近年来,了解 hMSC 的特性(例如它们在不同成熟阶段的电行为)变得更加重要。介电泳 (DEP) 是一种可以在非均匀电场中操纵细胞的方法,通过该方法可以获得有关细胞电特性的信息,例如细胞膜电容和介电常数。传统的 DEP 模式使用金属电极(例如三维电极)来表征细胞对 DEP 的响应。在本文中,我们提出了一种使用光导层构建的微流控装置,该装置能够通过充当具有易于顺应的几何形状的原位虚拟电极的光投影来操纵细胞。本文提出了一种用于表征 hMSCs 的方案,该方案展示了这种称为光诱导介电泳 (LiDEP) 的现象。我们表明,通过改变输入电压、光投影的波长范围和光源的强度等参数,可以优化 LiDEP 诱导的细胞响应,如细胞速度。将来,我们设想这个平台可以为无标记技术铺平道路,从而对 hMSCs 或其他干细胞系的异质群体进行实时特性分析。